ELISA was performed using a serial dilution of JMJD2B polyclonal antibody ( Cat # PAB14071 ). The wells were coated with the peptide used for immunisation of the rabbit. By plotting the absorbance against the antibody dilution, the titer of the crude serum was estimated to be 1 : 3,000.

WB using the antibody against JMJD2b diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

Determination of the titer To determine the titer, an ELISA was performed using a serial dilution of the antibody against human JMJD2b . The wells were coated with the peptide used for immunisation of the rabbit. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the antibody was estimated to be 1:3,000.

HeLa cells were stained with the antibody against JMJD2b and with DAPI. Cells were fixed with 4% formaldehyde for 10' and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the JMJD2b antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.